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Method for slicing heavy leather blade paraffin

A technology of paraffin slices and blades, which is applied in the direction of sampling devices, test samples preparation, etc., which can solve the problems of prolonging the soaking time of reagents, difficulties, and increased brittleness of materials

Inactive Publication Date: 2012-11-14
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thick leathery blades present considerable difficulties in making due to the exceptional thickness of the blades
First of all, it is difficult for the reagent to enter the tissue, but the goal cannot be achieved by simply prolonging the soaking time of the reagent, because the soaking time is too long, which will bring adverse effects to the material. For example, soaking in xylene for too long will increase the brittleness of the material , the tissue is fragile
Secondly, the dehydration process is not easy, and if the dehydration is not complete, hollow wax flakes will be formed. Therefore, the paraffin slices of thick leathery leaves often have broken materials, resulting in the failure of the slices.

Method used

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  • Method for slicing heavy leather blade paraffin
  • Method for slicing heavy leather blade paraffin
  • Method for slicing heavy leather blade paraffin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Specific steps are as follows:

[0037] 1) Take fresh and disease-free Cuban milk tree (Manilkara roxburghiana (Wight) Dubard) leaves (thickness 450-500 μm, leathery), use a single-sided blade to cut at 5-6 mm along both sides of the main vein, and cut the leaves with veins in the center The part is then cut transversely into small sections of 5-6mm.

[0038] 2) Fixation: The fixative is universal fixative (also known as FAA fixative, the formula is 90ml of 70% ethanol, 5ml of glacial acetic acid, and 5ml of formalin), the fixation time is 24h, and it can be stored for a long time.

[0039] 3) Dehydration: dehydration with ethanol, starting with 60% ethanol, increasing by 5%, each level is 4h, specifically 4h in 60% ethanol→4h in 65% ethanol→4h in 70% ethanol→4h in 75% ethanol→4h in 75% ethanol→80% ethanol 4h in 85% ethanol → 4h in 90% ethanol → 4h in 95% ethanol → 4h in 100% ethanol (the first time) → 4h in 100% ethanol (the second time).

[0040]4) Transparency: Tak...

Embodiment 2

[0050] 1) to 4) are as described in Example 1.

[0051] 5) Soft wax with a melting point of 52-54°C is used for wax penetration, and a solution of xylene: paraffin with a ratio of 1:1 (volume ratio) is used to transition to immerse in wax in a 37°C thermostat for 72 hours, and then replace it with pure paraffin in a 63°C thermostat Dip in wax, and change pure paraffin once or twice during the period, and then embed.

[0052] 6) As described in Example 1.

[0053] 7) Spreading: transition in 25% ethanol first, and then spread in warm water at a water temperature of 55°C.

[0054] 8) As described in Example 1.

[0055] 9) Bake the slices: bake the slices in an oven at 35°C for 36 hours.

[0056] 10) Dewaxing and rehydration: the procedure is specifically 100% ethanol: xylene in a 1:2 (volume ratio) solution for 1 min → 100% ethanol: xylene in a 1: 1 (volume ratio) solution for 1 min → 100% ethanol: Xylene in 2:1 (volume ratio) solution for 1min→100% ethanol 1min (first time)...

Embodiment 3

[0060] 1) to 6) are as described in Example 1.

[0061] 7) Spreading: transition in 35% ethanol first, and then spread in warm water at a water temperature of 48°C.

[0062] 8) As described in Example 1.

[0063] 9) Bake the slices: bake the slices in an oven at 35°C for 36 hours.

[0064] 10) Dewaxing and rehydration: the procedure is specifically 100% ethanol: xylene in a solution of 1:2 (volume ratio) for 2 minutes → 100% ethanol: xylene in a solution of 1:1 (volume ratio) for 2 minutes → 100% ethanol: Xylene is 2:1 (volume ratio) solution 2min→100% ethanol 2min (first time)→100% ethanol 2min (second time)→95% ethanol 2min→85% ethanol 2min→70% ethanol 2min→50 % ethanol 2min.

[0065] 11) Staining: the specific procedure is 1% safranin solution staining for 3 hours → 95% ethanol for 2 minutes → 100% ethanol for 2 minutes (the first time) → 100% ethanol for 2 minutes (the second time) → 100% ethanol: xylene is 2:1 2min in solution → 100% ethanol: xylene in 1:1 solution 2m...

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Abstract

The invention discloses a method for slicing heavy leather blade paraffin and relates to a method for slicing plant paraffin. Operating procedures include fixing, dehydrating, transparentizing, paraffin infiltration, embedding, slicing, dewaxing, dyeing and mounting. In the splicing process of the heavy leather blade paraffin, a fixing agent, a dehydrating procedure, a transparentizing procedure,a paraffin infiltration method, a dyeing method and mounting are improved, i.e., selection of the fixing agent, determination of the dehydrating procedure, determination of the transparentizing procedure, determination of the dewaxing procedure, the dyeing method and the mounting method are improved. The method is also suitable for other blades with different thicknesses and properties. By adopting the method, slice manufacturing of blade paraffin with different thicknesses and properties can be realized practically, and a uniform and practical manufacturing method for theoretical manufacturing of paraffin is popularized.

Description

technical field [0001] The invention relates to a plant paraffin sectioning method, in particular to a thick leathery leaf paraffin sectioning method. Background technique [0002] Paraffin sections are widely used in routine histological preparation techniques, mainly for observing the morphology and structure of cell tissues, and can be preserved for a long time. There are many procedures and procedures for making paraffin slices, and the cycle is long. Each link will affect the quality of slices. For example, problems such as slice bending and slice cracks are prone to occur in the process of slice production (Wu Minhua, Ke Zhanhua. Common problems and solutions in the process of paraffin slice production Method [J] Journal of Hainan Medical College, 2006, 12(3): 255). Although these problems can be overcome, if the material is broken, it cannot be remedied, and the section must be remade. The reason for material breakage can be traced back to many steps in the process o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/06G01N1/28G01N1/30
Inventor 刘育梅卢昌义
Owner XIAMEN UNIV
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